电化学疗法对乳腺癌MCF-7细胞凋亡及细胞内钙离子浓度的影响

Influence of electrochemical therapy on the apoptosis and intracellular Ca^(2+)concentration in breast cancer MCF-7 cells

目的探讨电化学疗法(ECT)对乳腺癌MCF-7细胞内钙离子浓度及细胞凋亡、坏死和细胞生长抑制的影响。方法将MCF-7细胞分为对照组及实验组,对照组为0 C(C:库仑),实验组按电量不同分为4组:3 C、5 C、10 C及20 C组。每组重复6孔,用不同电量处理MCF-7细胞后培养6 h和24 h,通过MTT法、GENMED细胞钙离子浓度比色法、流式细胞仪和激光共聚焦分别检测细胞生长抑制率、细胞内Ca2+浓度、观察凋亡和坏死情况。组间均数比较采用方差分析,应用SNK方法进行两两比较,应用5×2析因分析探讨电量与时间对细胞的影响。结果不同电量干预MCF-7细胞后培养6 h及24 h,细胞生长抑制率(6 h:F=508.43,P=0.000;24 h:F=232.76,P=0.000)、坏死率(6 h:F=2282.07,P=0.000;24 h:F=2644.60,P=0.000)、细胞内Ca2+浓度(6 h:F=1416.06,P=0.000;24 h:F=2394.26,P=0.000)随电量增加而增加,电量及培养时间之间具有交互作用(F=288.93,P=0.000;F=2305.39,P=0.000;F=1651.96,P=0.000)。流式细胞仪检测细胞凋亡率结果显示,细胞凋亡率在3、5、10 C组明显增高,而20 C组细胞凋亡率降低,电量及培养时间有交互作用(F=51.20,P=0.000)。结论 ECT可抑制乳腺癌MCF-7细胞株的生长并诱导其凋亡,并引起细胞内Ca2+浓度升高。低剂量ECT主要诱导细胞凋亡,高剂量ECT主要引起细胞坏死。

Objective To investigate the influence of electrochemical therapy （ECT） on intracellular Ca^2＋ concentration, apoptosis and necrosis, and proliferation inhibition in breast cancer MCF-7 cells. Methods MCF-7 cells were randomly divided into 5 groups treated by different quantities of electricity,i, e. , 0 e（control group） ,3 C,5 C,10 C,20 C （experimental groups）. We replicated 6 wells in each group. The cells were cultured for 6 h and 24 h after electrochemical treatment. MTT method, GENMED calcium colorimetric assay and flow cytometry assay were used to detect the inhibitory rate of cells growth and intracellular Ca^2＋ concentration, respectively. The confoeal laser scanning microscope was used to observe the apoptosis and necrosis of MCF-7 ceils. The means between the two groups were compared using analysis of variance, SNK method was used for pairwise comparison, and 5 × 2 factorial analysis was performed to explore the effect of quantity of electricity and time on the cells. Results At 6 h and 24 h of culture after ECT with different quantities of electricity, the inhibition rate of MCF-7 cells （6h： F = 508.43, P = 0.000; 24 h： F = 232.76, P = 0. 000 ）, necrosis rate （6 h：F=2282.07,P=0.000;24 h：F=2644.6,P=0. 000）,and intracellular Ca^2＋ concentration （6 h：F= 1416.06, P = 0.000; 24 h： F = 2394.26, P = 0.000 ） were significantly increased with the increase of quantity of electricity,and the interaction was observed between quantity of electricity and culture time （F = 288.93, P = 0. 000 ; F= 2305.39, P = 0. 000 ; F = 1651.96, P = 0. 000 ）. Flow cytometry showed that the apoptosis rate was significantly increased in 3 C,5 C, 10 C groups, and then decreased in 20 C group, the interaction was observed between quantity of electricity and culture time （F= 51.20,P= 0. 000）. Conclusions ECT can inhibit the cell growth and induce the apoptosis of MCF-7 cells and increase the concentration of intracellular Ca^2＋. However,low dose of ECT mainly induces cell apoptosis and high dose of ECT mainly causes cell necrosis.